抗菌肽Spinigerin α基因定点突变体的构建

Construction of the site-directed mutants for antibacterial peptide Spinigerin

通过对Spinigerin α抗菌肽的结构分析,结合抗菌肽的抗菌机理,根据毕赤酵母的偏爱密码子对该抗菌肽进行定点突变。将亲水性氮端的碱性赖氨酸分别突变为碱性更强的精氨酸、酸性的天冬氨酸以及中性的丙氨酸,将疏水性碳端的中性亮氨酸突变成碱性的精氨酸、酸性的天冬氨酸。将定点突变后的Spinigerin α基因按正确阅读框克隆至穿梭质粒pPICZ α-A上,经PCR鉴定、序列分析,所转化的宿主酵母GS115中含有该突变体,结果表明已成功构建了抗菌肽Spinigerin α基因的突变体,为获得高效的抗菌效果的抗菌肽奠定一定的基础。

The structural analysis of antimicrobial peptides Spinigerin a, combined with the antibacterial mechanism of antimicrobial peptides, using the yeast Pichia preferred codons of the antimicrobial peptides conduct site-directed mutagenesis. The N-terminal （hydrophilic side） alkaline lysine were to mutate into the more stronger alkaline arginine, acidic aspartic acid and neutral alanine; the C-terminal （hydrophobic side） of the neutral leucine were to mutate into the alkaline arginine and acidic aspartate. Site-directed mutagenesis Spinigerin a gene in the correct reading frame was cloned into shuttle plasmid pPICZa-A, was identified by PCR, sequence analysis, the transformed yeast host pastoris GSl15 containing the mutant, the result was successfully constructed antimicrobial peptides Spinigerin-a gene mutants, and lay a foundation for obtain efficient antibacterial effect of antimicrobial peptides.